 |
PDBsum entry 5ifr
|
|
|
|
References listed in PDB file
|
 |
|
Key reference
|
|
|
Title
|
|
A cascading activity-Based probe sequentially targets e1-E2-E3 ubiquitin enzymes.
|
|
|
Authors
|
|
M.P.Mulder,
K.Witting,
I.Berlin,
J.N.Pruneda,
K.P.Wu,
J.G.Chang,
R.Merkx,
J.Bialas,
M.Groettrup,
A.C.Vertegaal,
B.A.Schulman,
D.Komander,
J.Neefjes,
F.El oualid,
H.Ovaa.
|
|
|
Ref.
|
|
Nat Chem Biol, 2016,
12,
523-530.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
Abstract
|
|
|
Post-translational modifications of proteins with ubiquitin (Ub) and
ubiquitin-like modifiers (Ubls), orchestrated by a cascade of specialized E1, E2
and E3 enzymes, control a wide range of cellular processes. To monitor catalysis
along these complex reaction pathways, we developed a cascading activity-based
probe, UbDha. Similarly to the native Ub, upon ATP-dependent activation by the
E1, UbDha can travel downstream to the E2 (and subsequently E3) enzymes through
sequential trans-thioesterifications. Unlike the native Ub, at each step along
the cascade, UbDha has the option to react irreversibly with active site
cysteine residues of target enzymes, thus enabling their detection. We show that
our cascading probe 'hops' and 'traps' catalytically active Ub-modifying enzymes
(but not their substrates) by a mechanism diversifiable to Ubls. Our founder
methodology, amenable to structural studies, proteome-wide profiling and
monitoring of enzymatic activity in living cells, presents novel and versatile
tools to interrogate Ub and Ubl cascades.
|
|
|
|
|
|
|
